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InsTAclone™ PCR Cloning Kit*
* Available in certain countries only.

#K1213	for 10 reactions
#K1214	for 30 reactions
Related Documents (in pdf, ~50-300 KB): Detailed Protocol: #K1213, #K1214 MSDS (English) MSDS (English-USA) MSDS (German) Flyer

 

Features

• Fast cloning - approximately one hour from PCR completion to cell plating.
• High efficiency - more than 90% of the recombinant clones contain the target DNA.
• One-step procedure - additional modifications of PCR fragment are not required.
• Compatibility - compatible with Taq, Tth, Tfl and other non-proofreading DNA polymerases, as well as with the Fermentas Long PCR Enzyme Mix.
• Convenience of pTZ57R/T cloning vector:
  • ready-to-use: linearized and 3'-ddT tailed,
  • MCS designed for easy manipulation with a cloned insert,
  • blue/white screening,
  • M13/pUC primer sites for sequencing or PCR screening,
  • T7 promoter for in vitro transcription of the insert.

TransformAid™ Bacterial Transformation Kit. InsTAclone™ PCR Cloning Kit, flyer in pdf, 319 KB

Description
The InsTAclone™ PCR Cloning Kit is a TA system for direct one-step cloning of PCR products with 3'-dA overhangs (1). Overhangs of this type are generated by Taq DNA polymerase and other thermostable DNA polymerases which lack proofreading activity. The kit includes a ready-to-use cloning vector pTZ57R/T (see Fig.2) constructed on the backbone of the pTZ57R vector. The 3'-ddT overhangs at both ends of the cloning site prevent recircularization of the vector during ligation, therefore resulting in high cloning yields and low background. To increase the speed, convenience and efficiency of your cloning reaction, the InsTAclone™ PCR Cloning Kit has been combined with the unique TransformAid™ Bacterial Transformation Kit**. According to our protocol, ligation and preparation of competent cells is performed in parallel. Therefore, it only takes approximately one hour from the completion of PCR to cell plating. Our transformation protocol is often faster than transformation of commercially available competent cells.

Application
TA cloning.

Quality Control
The kit is tested by the resultant cloning efficiency of a control PCR product. Typical yield of the recombinant clones is higher than 90%.

Components of the Kit

• Vector pTZ57R/T
• T4 DNA Ligase
• 5X Ligation Buffer
• Control PCR Fragment
• Control DNA (pTZ57R with insert)
• Vector pTZ57R DNA (without insert)
• Water, nuclease-free
• TransformAid™ Bacterial Transformation System:
  • C-Medium
  • T-Solution (A)
  • T-Solution (B)
• Detailed Protocol (in pdf, ~91 KB):
  • #K1213, #K1214

Note
E.coli strains are not included.
* Lyophilized bacterial strains GM2163 (#M0099) and JM107 (#M0109) can be provided free of charge upon request, with any purchase.
** The TransformAid™ Bacterial Transformation Kit can be used with most E.coli strains commonly used for cloning, including JM107, JM109, XL1-Blue, SURE, TOPP2, W3110, NM527, AD494, CJ236, GM2163, C600, BL21.

Figure 1. PCR product cloning procedure.

Figure 2. Restriction map of vector pTZ57R/T.

Related Products

Taq DNA Polymerase (recombinant) Taq DNA Polymerase (native) Long PCR Enzyme Mix dNTP Mixes dNTP Set Ampicillin E.coli FastMedia™ IPTG, dioxane free IPTG Solution, ready-to-use X-Gal X-Gal Solution, ready-to-use Sequencing Primers: M13/pUC Primers T7 Promoter Sequencing Primer, 20-mer T7 RNA Polymerase T7 Transcription Kit GeneJET™ Plasmid Miniprep Kit DNA Extraction Kit FastDigest® Restriction Enzymes E.coli strains GM2163, JM107*

Reference

1. Clark, J.M., Novel non-templated nucleotide addition reactions catalyzedby procaryotic and eucaryotic DNA polymerases, Nucl. Acids Res., 16, 9677-9686, 1988.

Updated kovo 04, 2008 10:20